Current progress and future prospect of microalgal biomass harvest using various flocculation technologies.

Microalgae have been extensively studied for the production of various valuable products. Application of microalgae for the production of renewable energy has also received increasing attention in recent years. However, high cost of microalgal biomass harvesting is one of the bottlenecks for commercialization of microalgae-based industrial processes. Considering harvesting efficiency, operation economics and technological feasibility, flocculation is a superior method to harvest microalgae from mass culture. In this article, the latest progress of various microalgal cell harvesting methods via flocculation is reviewed with the emphasis on the current progress and prospect in environmentally friendly bio-based flocculation. Harvesting microalgae through bio-based flocculation is a promising component of the low-cost microalgal biomass production technology.

Characterization of the flocculating agent from the spontaneously flocculating microalga Chlorella vulgaris JSC-7.

High cost of biomass recovery is one of the bottlenecks for developing cost-effective processes with microalgae, particularly for the production of biofuels and bio-based chemicals through biorefinery, and microalgal biomass recovery through cell flocculation is a promising strategy. Some microalgae are naturally flocculated whose cells can be harvested by simple sedimentation. However, studies on the flocculating agents synthesized by microalgae cells are still very limited. In this work, the cell flocculation of a spontaneously flocculating microalga Chlorella vulgaris JSC-7 was studied, and the flocculating agent was identified to be cell wall polysaccharides whose crude extract supplemented at low dosage of 0.5 mg/L initiated the more than 80% flocculating rate of freely suspended microalgae C. vulgaris CNW11 and Scenedesmus obliquus FSP. Fourier transform infrared (FTIR) analysis revealed a characteristic absorption band at 1238 cm(-1), which might arise from PO asymmetric stretching vibration of [Formula: see text] phosphodiester. The unique cell wall-associated polysaccharide with molecular weight of 9.86×10(3) g/mol, and the monomers consist of glucose, mannose and galactose with a molecular ratio of 5:5:2. This is the first time to our knowledge that the flocculating agent from C. vulgaris has been characterized, which could provide basis for understanding the cell flocculation of microalgae and breeding of novel flocculating microalgae for cost-effective biomass harvest.

Characterization of flocculating agent from the self-flocculating microalga Scenedesmus obliquus AS-6-1 for efficient biomass harvest.

In the present work, the extracellular biopolymers from the self-flocculating microalga Scenedesmus obliquus AS-6-1 were studied. It was revealed that the self-flocculation of the microalgal cells was mediated by cell wall-associated polysaccharides with a molecular weight of 127.9 kDa. Sugar compositions analysis indicated that the monomers consist of glucose, mannose, galatose, rhamnose and fructose with the molar ratio of 8:5:3:2:1. Addition of 0.6 mg/L purified flocculating agent resulted in the fast flocculation of freely suspended cells of S. obliquus and Chlorella vulgaris. The flocculating activity is stable between pH 6 and 8 and at 20-60°C.

Bioflocculant production from Solibacillus silvestris W01 and its application in cost-effective harvest of marine microalga Nannochloropsis oceanica by flocculation.

Microalgae are widely studied for biofuel production, however, current technologies to harvest microalgae for this purpose are not well developed. In this work, a bacterial strain W01 was isolated from activated sludge and identified as Solibacillus silvestris. Bioflocculant in the culture broth of W01 showed 90% flocculating efficiency on marine microalga Nannochloropsis oceanica, and no metal ion was required for the flocculation process. Chemical analysis of the purified bioflocculant indicated that it is a proteoglycan composed of 75.1% carbohydrate and 24.9% protein (w/w). The bioflocculant exhibits no effect on the growth of microalgal cells and can be reused to for economical harvesting of N. oceanica. This is the first report that strain of S. silvestris can produce bioflocculant for microalgae harvest. The novel bioflocculant produced by W01 has the potential to harvest marine microalgae for cost-effective production of microalgal bioproducts.

Establishment of an efficient genetic transformation system in Scenedesmus obliquus.

Scenedesmus obliquus belongs to green microalgae, which is attracting attention as a feedstock for biofuels production and biorefinery as well as in bioremediation of environmental pollutants, making its genetic modifications for more efficient growth and accumulation of aimed metabolites significant. However, the genetic transformation system of S. obliquus is still not well established. In the current work, S. obliquus was transformed via electroporation using a plasmid containing chloramphenicol resistance gene (CAT) as a selectable marker and the green fluorescent protein gene (gfp) as a reporter. Using the optimized transformation conditions, the transformation efficiency was 494±48 positive transgenic clones per 10(6) recipient cells, which is more efficient comparing with those reported in other microalgal transformation studies. Green fluorescence was observed after six months of cultivation, and CAT-specific products were also detected in the transformants by PCR, Southern blot and RT-PCR analysis. This is the first report on establishing such an efficient and stable transformation system for S. obliquus, a prerequisite for both functional genomic studies and strain improvement for other biotechnology applications of this important microalgal species.

Role of nsdA in negative regulation of antibiotic production and morphological differentiation in Streptomyces bingchengensis.

To investigate the function of nsdA in Streptomyces bingchengensis, it was cloned and sequenced, which presented an 89.89% identity with that of S. coelicolor. The lambdaRED-mediated PCR-targeting technique was used to create nsdA replacement in the S. bingchengensis_226541 chromosome. The nsdA disruption mutant, BC29, was obtained, which produced more pigment and spores than did the ancestral strain. HPLC analysis revealed that the disruption of nsdA efficiently increased milbemycin A(4) production and nanchangmycin production by 1.5-fold and 9-fold, respectively. Complementation of the nsdA mutation restored the phenotype and antibiotic production. These results showed that nsdA negatively affected sporulation and antibiotic production in S. bingchengensis.